Geriet al.,Science 367 , 1091–1097 (2020) 6 March 2020 5of7
Fig. 4. Intra- and extrasynapticmmapping within a two-cell system.
(A) Schematic depicting Jurkat (CD45RO+/PD-1+/CD3+) and JY (PD-L1+/CD19+)
two-cell system enhanced by the presence of staphylococcal enterotoxin D (SED)
for antibody targeting of intrasynaptic PD-L1 (expressed on JY cells) and
extrasynaptic CD45RO (distinctly expressed on Jurkat cells) for selective trans or
cis labeling. (B) Flow cytometry analysis of antibody targeting of PD-L1 on JY
PD-L1 B cells withmMap by using 10-min light irradiation (left) shows both cis-
and trans-cellular labeling. Peroxidase-based targeted labeling for 0.5 min (right)
results in nearly complete cis- and trans-cellular labeling. (C) Flow cytometry
analysis of antibody targeting of CD45RO on Jurkat T cells withmMap by using
10-min light irradiation (left) shows only cis-cellular labeling. Peroxidase-based
targeted labeling for 0.5 min (right) results in nearly complete cis- and trans-
cellular labeling. (D) Two-cell system confocal microscopy images of isotype-
targeted (10-min light irradiation) or PD-L1–targeted (0.5-, 2-, or 10-min light
irradiation) by usingmMap (left) or isotype-targeted (1 min) or PD-L1–targeted
(0.5 or 1 min) by using peroxidase-based labeling (right). Cells were imaged for
biotinylation (green), CD3 surface expression (magenta), and nuclei (blue). Scale
bar, 5mm. Duplicate images are shown below each condition.RESEARCH | RESEARCH ARTICLE